一株地黄根腐病病原菌的分离鉴定及生物学特性研究

doi:10.13560/j.cnki.biotech.bull.1985.2019-0771

生物技术通报 ›› 2020, Vol. 36 ›› Issue (1): 37-44.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0771

一株地黄根腐病病原菌的分离鉴定及生物学特性研究

王亚利¹, 康春晓¹, 杨传臻¹, 魏艺璇¹, 王瑞飞^1,2, 李明军^1,3, 杨清香^1,2

1. 河南师范大学生命科学学院,新乡 453007;
2. 资源微生物与功能分子河南省高校重点实验室培育基地（河南师范大学）,新乡,453007;
3. 河南省道地药材保育及利用工程技术研究中心/绿色药材生物技术河南省工程实验室,新乡 453007

收稿日期:2019-08-24 出版日期:2020-01-26 发布日期:2020-01-08
作者简介:王亚利,硕士研究生,研究方向：资源与微生物学;E-mail：17797700943@163.com
基金资助:
国家自然科学基金项目（21277041,21477035,U1504301）,河南省高等学校重点科研项目（15A180016）

Isolation and Identification of a Root Rot Pathogen of Rehmannia glutinosa and Its Characterization

WANG Ya-li¹, KANG Chun-xiao¹, YANG Chuan-zhen¹, WEI Yi-xuan¹, WANG Rui-fei^1,2, LI Ming-jun^1,3, YANG Qing-xiang^1,2

1. College of Life Sciences,Henan Normal University,Xinxiang 453007;
2. Key Laboratory for Microorganisms and Functional Molecules（Henan Normal University）,University of Henan Province,Xinxiang 453007;
3. Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs in Henan Province,Xinxiang 453007

Received:2019-08-24 Published:2020-01-26 Online:2020-01-08

摘要/Abstract

摘要： 分离鉴定地黄根腐病的病原菌,对其致病性和生物学特性进行研究。利用平板分离法对地黄根腐病的病原菌进行分离纯化;利用测序技术和遗传进化分析对所得菌株进行鉴定;利用回接试验对所得菌株进行致病性检测;利用生长监测,确定所得菌株的各项生理学特性。获得一株菌落橙黄色,边缘整齐,表面光滑、湿润菌体椭圆形,有疑似出芽现象的真菌;该菌与帕鲁迪根红酵母（Rhodotorula paludigena）簇集在一起,置信度为91,遗传距离最近,具有密切的亲缘关系;该酵母菌接种侵染20 d后,地黄块根根腐病发病率达到100%;该菌株最适生长温度为28℃,最适生长pH为5.0;该酵母菌株能耐受阿魏酸、香草酸和对羟基苯甲酸（与根腐病发病密切相关的地黄根部分泌物）的最高浓度为0.5 g/L。初步证实了Rhodotorula paludigena DH5能够引起地黄根腐病,且该病原在富含酚酸类化合物的地黄根际具较强的生态适应能力。

关键词: 地黄, 根腐病, 酵母菌, 酚酸, 生物学特性

Abstract: We aimed to isolate and identify the pathogens resulting in Rehmannia glutinosa root rot as well as study its pathogenicity and biological characteristics. In this study,streak plate method was used to isolate and purify the pathogens of rot root in R. glutinosa. ITS sequencing and genetic evolution analysis were used to identify the isolated strains. Inoculation experiment was used to confirm the pathogenic of the isolated potential pathogens. The growth test was performed to analyze physiological characteristics of the isolated stain. One of the obtained strains presented the colony morphology of orange color,the neat edge,the smooth and moist surface,and microscopic morphology with oval shape,suspected to be a fungus with buds. The strain was clustered together with the Rhodotorula paludigena at confidence of 91 and they displayed the close genetic relation. R. glutinosa roots inoculated with the strain and the incidence was 100% at day 20. The optimal growth temperature and pH of the strain was 28℃ and 5.0,respectively. The strain grew well in the medium with 0.5 g/L ferulic acid,vanillic acid and p-hydroxybenzoic acid,i.e.,the secretion in the root of R. glutinosa that was closely related to the root rot of R. glutinosa. It is preliminarily verified that R. paludigena DH5 may cause the root rot of R. glutinosa and well adapts to the rhizosphere environment containing a large content of phenolic compounds.

Key words: Rehamannia glutinosa, phytophthora root rot, Rhodotorula paludigena, phenolic acid, biological characteristics

王亚利, 康春晓, 杨传臻, 魏艺璇, 王瑞飞, 李明军, 杨清香. 一株地黄根腐病病原菌的分离鉴定及生物学特性研究[J]. 生物技术通报, 2020, 36(1): 37-44.

WANG Ya-li, KANG Chun-xiao, YANG Chuan-zhen, WEI Yi-xuan, WANG Rui-fei, LI Ming-jun, YANG Qing-xiang. Isolation and Identification of a Root Rot Pathogen of Rehmannia glutinosa and Its Characterization[J]. Biotechnology Bulletin, 2020, 36(1): 37-44.

参考文献

[1] Yang Q, Wang R, Xu Y, et al.Dynamic change of the rhizosphere microbial community in response to growth stages of consecutively monocultured Rehmanniae glutinosa[J]. Biologia, 2016, 71(12):1320-1329.
[2] Jung DL, Song JY, ILL MC, et al.Resveratrol synthase transgene expression and accumulation of resveratrolglycoside in Rehmannia glutinosa[J]. Molecular Breeding, 2005, 16:219-233.
[3] Wu L, Wang J, Huang W, et al.Plant-microbe rhizosphere interactions mediated by Rehmannia glutinosa root exudates under consecutive monoculture[J]. Scientific Reports, 2016, 5:15871.
[4] 黄艳琴. 地黄常见病害的发生与防治[J]. 河南农业, 2006(8):22-22.
[5] Bååth E, Anderson T.Comparison of soil fungal/bacterial ratios in a pH gradient using physiological and PLFA-based techniques[J]. Soil Biology & Biochemistry, 2003, 35(7):955-963.
[6] Lauber C, Hamady M, Knight R, et al.Pyrosequencing-based assessment of soil pH as a predictor of soil bacterial community structure at the continental scale[J]. Applied & Environmental Microbiology, 2009, 75(15):5111.
[7] Aciego C, Brookes P.Substrate inputs and pH as factors controlling microbial biomass, activity and community structure in an arable soil[J]. Soil Biology & Biochemistry, 2009, 41(7):1396-1405.
[8] Bengtson P, Sterngren A, Rousk J.Archaeal abundance across a pH gradient in an arable soil and its relationship to bacterial and fungal growth rates[J]. Applied & Environmental Microbiology, 2012, 78(16):5906-5911.
[9] Lipson D, Schmidt S.Seasonal changes in an alpine soil bacterial community in the colorado rocky mountains.[J]. Applied & Environmental Microbiology, 2004, 70(5):2867.
[10] Nemergut D, Costello E, Meyer A, et al.Structure and function of alpine and arctic soil microbial communities.[J]. Research in Microbiology, 2005, 156(7):775-784.
[11] 国春菲. 土壤盐分和pH对滨海盐土土壤微生物多样性的影响[D]. 杭州:浙江农林大学, 2013.
[12] 李振方, 杨燕秋, 谢冬凤, 等. 连作条件下地黄药用品质及土壤微生态特性分析[J]. 中国生态农业学报, 2012, 20(2):217-224.
[13] 杜家方. 地黄(Rehmannia glutinosa Libosch. )根区土壤中酚酸类物质的化感作用研究[D]. 郑州:河南农业大学, 2009.
[14] Qu X, Wang J.Effect of amendments with different phenolic acids on soil microbial biomass, activity, and community diversity[J]. Applied Soil Ecology, 2008, 39(2):172-179.
[15] 吴林坤, 黄伟民, 王娟英, 等. 不同连作年限野生地黄根际土壤微生物群落多样性分析[J]. 作物学报, 2015, 41(2):308-317.
[16] Kefeli V, Kalevitch V, Borsari B.Phenolic cycle in plants andenvi-ronment[J]. J Cell & Mol Biol, 2003, 2(1):13-18.
[17] Chan Y.Utilization of simple phenolics for dinitrogen fixation by soil diazotrophic bacteria[J]. Plant and Soil, 1986, 90(1/2/3):141-150.
[18] Wang R, Wang Y, Yang Q, et al.Unraveling the characteristics of the microbial community and potential pathogens in the rhizosphere soil of Rehmannia glutinosa with root rot disease[J]. Applied Soil Ecology, 2018, doi:org/10.1016/japsoil.2018.07.001.
[19] 桓明辉, 李杨, 刘晓辉, 等. 1株产纤维素酶木霉菌种的鉴定[J]. 微生物学杂志, 2012, 32(5):65-68.
[20] Zhao Z, Gao X, Yang D, et al.Field detection of canker-causing bacteria on kiwifruit trees:Pseudomonas syringae, pv. actinidiae, is the major causal agent[J]. Crop Protection, 2015, 75:55-62.
[21] 廖长宏, 陈军文, 吕婉婉, 等. 根茎类药用植物根腐病研究进展[J]. 中药材, 2017, 40(2):493-498.
[22] 王立新, 刘正坪. 黄芪根腐病病原菌鉴定[J]. 华北农学报, 1994, 9(2):107-109.
[23] 罗文富, 喻盛甫, 贺承福, 等. 三七根腐病病原及复合侵染的研究[J]. 植物病理学报, 1997, 27(1):85-91.
[24] 沈清清, 刘芳, 胡彦. 药用植物根腐病病原菌研究进展[J]. 北方园艺, 2014, 11:187-190.
[25] Song H, Gao XL, Wang XM, et al.Effects of exogenous phthalic and cinnamic acids on enzyme activities and microbial community structures in rhizosphere soil of Adzuki Bean.[J]. Agricultural Science & Technology, 2017(10):1935-1940.
[26] 刘苹, 高新昊, 孙明, 等. 3种酚酸类物质对花生发芽和土壤微生物的互作效应研究[J]. 江西农业学报, 2012, 24(8).
[27] 母容, 潘开文, 王进闯, 等. 阿魏酸、对羟基苯甲酸及其混合液对土壤氮及相关微生物的影响[J]. 生态学报, 2011, 31(3):793-800.
[28] Inderjit, Streibig JC, Olofsdotter M. Joint action of phenolic acid mixtures and its significance in allelopathy research[J]. Physiologia Plantarum, 2010, 114(3):422-428.

一株地黄根腐病病原菌的分离鉴定及生物学特性研究

Isolation and Identification of a Root Rot Pathogen of Rehmannia glutinosa and Its Characterization

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	杨东亚, 祁瑞雪, 李昭轩, 林薇, 马慧, 张雪艳. 黄瓜茄病镰刀菌拮抗芽孢杆菌的筛选、鉴定及促生效果[J]. 生物技术通报, 2023, 39(2): 211-220.
[2]	文畅, 刘晨, 卢诗韵, 许忠兵, 艾超凡, 廖汉鹏, 周顺桂. 一株新的多重耐药福氏志贺菌噬菌体生物学特性及基因组分析[J]. 生物技术通报, 2022, 38(9): 127-135.
[3]	王春艳, 腊贵晓, 苏秀红, 李萌, 董诚明. 地黄不同时期内生促生细菌的筛选及其促生特性分析[J]. 生物技术通报, 2022, 38(4): 242-252.
[4]	杨瑞先, 刘萍, 王祖华, 阮宝硕, 汪智达. 牡丹根腐病原菌拮抗细菌抑菌活性物质分析[J]. 生物技术通报, 2022, 38(2): 57-66.
[5]	赵林艳, 官会林, 向萍, 李泽诚, 柏雨龙, 宋洪川, 孙世中, 徐武美. 白及根腐病植株根际土壤微生物群落组成特征分析[J]. 生物技术通报, 2022, 38(2): 67-74.
[6]	高惠惠, 贾晨波, 韩琴, 苏建宇, 徐春燕. 宁杞7号枸杞根腐病发生的微生物学机制[J]. 生物技术通报, 2022, 38(12): 244-251.
[7]	黄景晓, 尚俊康, 陈慧敏, 沈嘉旻, 黎圆圆, 喻玉立, 倪进东, 林伯坤. 一株烈性沙门氏菌噬菌体的生物学特性及基因组分析[J]. 生物技术通报, 2021, 37(6): 136-146.
[8]	张雅静, 宋美燕, 张怡静, 房庆, 杨俊, 彭德良, 黄文坤, 彭焕, 朱英波, 孔令安. 兼防黄瓜根腐病和根结线虫病的淡紫拟青霉和哈茨木霉的筛选[J]. 生物技术通报, 2021, 37(2): 40-50.
[9]	吕燕, 王文彬, 苟琪, 王英娜, 李靖宇, 刘建利. 根腐病对宁夏枸杞根区土壤丛枝菌根真菌群落的影响[J]. 生物技术通报, 2021, 37(12): 29-40.
[10]	要雅倩, 成娜娜, 李培根, 卢毅, 王彦刚, 林榕姗, 周波, 王冰. 解淀粉芽胞杆菌Bacillus amyloliquefaciens T-6的分离鉴定及抗病促生潜力[J]. 生物技术通报, 2020, 36(9): 202-210.
[11]	杜建峰, 吴伟, 张晓英, 李洋, 丁新华. 番茄颈腐根腐病的发生及其防治研究进展[J]. 生物技术通报, 2020, 36(10): 200-206.
[12]	赵晓霞, 牛世全, 文娜, 苏锋锋. 黄芪根腐病生防芽孢杆菌的筛选鉴定与盆栽防效试验[J]. 生物技术通报, 2019, 35(9): 107-111.
[13]	罗冬章, 罗惠娜, 詹小舒, 李心怡, 黄曼晴, 林洁微, 黄绮亮, 洪纯, 林好美, 陈胜锋, 王丙云. 猫4种不同来源间充质干细胞的生物学特性比较[J]. 生物技术通报, 2019, 35(7): 39-45.
[14]	郭萌萌, 周延清, 段红英, 杨珂, 邵露营. 基于地黄转录组数据的SNP标记开发与地黄指纹图谱构建[J]. 生物技术通报, 2019, 35(11): 224-230.
[15]	许艳俊, 李静媛. pH和Ca2+协同作用对酵母代谢及细胞膜功能的影响[J]. 生物技术通报, 2018, 34(3): 208-216.